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I. INTRODUCTION
Buchanaia lanzan (Chironji) belongs to family Anacardiaceae is depleting with a very fast rate and
presently categorized under the threatened species (registered in IUCN list), Therefore, conservation and
sustainable use of this type of species is an important necessity for ecologically sustainable development,
food security and development of socio-economically poor communities of the nation. These species play
very important role in the socio-economic condition of tribal population of the state. (Genetic Resources of
Tropical Underutilized Fruits in India (Malik, et al., 2010). Chironji is a multipurpose tree, provides food,
fodder, timber, medicines and gives monitory reward to tribal community of India. It possesses highly
medicinal value like Chironji kernel paste is an excellent skin conditioner and used as ointment for various
skin diseases. In addition to this, its leaves are reported to be valued for their tonic and cardio tonic
properties and their powder is a common medicine for wounds. The roots are acrid, astringent, cooling,
depurative and constipating, and are useful in treatment of diarrhea. No systematic study has been
conducted to identify good cultivars or selection of elite clones in this important minor fruit throughout the
country especially in the state of Chhattisgarh. Thus there is an urgent need to identify and characterize
superior selections in Chironji for promotion of this highly potential indigenous horticultural fruit crop in
Chhattisgarh.
Chironji has alternate bearing nature as Mango. It is highly heterozygous plant, which is cross
pollinated contributes genetic variation. Characterization of morphological traits is also essential to
document and study the genetic diversity within the collection, which can facilitate the establishment of core
collections that fully represent the diversity of the genotypes. At present this plant is grouped as an
underexploited and non-nationalized minor forest produce so it is free for collection. Selection of all the
wild genotypes for each regionbased on the information elicited from routine questioning and advice by
experts and local inhabitants who are involved in collection and selling of minor forest based produce;
especially Chironji, one of the important multipurpose forest species brings income to them. Fruit size, Fruit
colour, Seed weight Seed Purity, Seed moisture content, Germination percentage are the aspects on which
analysis was based. In present study, Exploration, Collection & Characterization of genetic diversity of
Chironji through use of the morphological analysis from Chhattisgarh was studied on the basis of seed.
Methods and materials:
1. Study area evaluation for morphological traits
The study was conducted in three major Agro climatic zones of Chhattisgarh. Total of 27 wild
Chironji genotypes with three replicates of nine trees with maximally 15km distance from each other were
referred to as a population. Malvolti et al., (1994), from Chhattisgarh plains zone, Bastar plateau Zone and
Northern Hill Zones were marked for detailed study.
Methodology:
1. Fruits collection:
Developing ripened and well mature fruits from each of selected tree were collected at 15 days
Interval. Fruit collection was done during the period of May (second fortnight) to June (first fortnight) in the
year 2014 and 2015. Fruits were generally collected from the ground.
2. Seed processing and samples preparation:
Collected fruits were brought to the laboratory and soaked in cold water for four days to remove the
pulp. Dried nuts locally called ‘Guthali’ were broke manually.
3. Morphological attributes
The main purposes of seed testing are to:
Extract from ‘Guide to Handling of Tropical and Subtropical Forest Seed’ by Lars Schmidt, Danida
Forest Seed Centre. 2000.
1. Seed Weight :
Seed weight refers the weight of 1,000 seeds according to ISTA. However, in the market, seed
weight refers to the amount seed per kilogram.
1. 100 seeds for 8 replicates were weighed separately, average weight of 100 seeds were calculated,
and multiply by 10
1000 seeds weight = 100 seeds weight x 10
2. The calculation accuracy was determine through statistic analysis using the formula below
2. Seed Purity:
Purity is the composition by weight of pure seed in a sample. Seed samples must be carefully
prepared in order to represent the whole seed lot. Sub-samples should be drawn from 2-3 different places
within the seed lot, mixed thoroughly, and then divided into test samples of about 2,500 seeds.
1. The lot of seed samples were weighed
2. The pure from impure seed was separated, using the following criteria listed in Table.
Seed Purity Criteria
Pure seed Impure Seed
Mature seed Seeds of other species
Undamaged seed Stones ,leaves, twigs etc
Undersized seed Seed wings
Immature seeds Pieces of seed less than half original size
Pieces of seed more than half original size Legume seed without seed coat
Shriveled seeds
Germinated seeds
Weigh the pure fraction and calculate to a percentage using the formula below.
3. Moisture Content: Moisture content represents the percentage of water in seed, determined through
two methods.
1) Direct method / Oven method:
2) Indirect method
Methodology of direct method
1. 2 empty containers were weighted (at least 5g for small seed, and at least 10g for large seed).
2. 10 gm of seeds were taken into small pieces. The weight of the container was not included.
3. 10 gm of seed were dried in oven at ± 103°C for ± 17 hours.
4. The samples allowed cool in an incubator for 15 minutes
5. The dry sample weighted and the moisture content was calculated as follows:
To ensure accuracy, statistical analysis has determined that the largest difference between replicates
is as illustrated below.
Average Moisture content
1000 seed weight
< 12% 12%-25% >25%
4. Seed Viability:
As per ISTA protocol viability is tested through;
1) Cutting test –
1. Sample of 100 randomly collected seeds were prepared
2. Each seed was cuted along
3. Fresh embryo part was regarded as viable.
Calculation of seed viability
freshseeds
Viability =
Totalcutseeds
5. Germination tests:
To determine viability under optimum conditions, where optimal conditions refers to best pre-
treatment; sufficient moisture; optimum temperature (about 25°C), and most suitable medium (without pest
and fungi infection).
1. Seed samples, each with 100 seeds were prepared for each pretreatment.
2. Raised beds of 10m x 1m x 0.15m size were prepared and well decomposed FMY was and NPK
properly mixed into top soil of the bed
3. Each raised bed consisted of five parts for five Pretreatments and in each plot 100 seeds. The seeds
were sown 2-3 cm deep in the nursery bed.
4. The germinated seeds were counted and stop when germination in all replicates.
5. The observations were recorded as germination percentage, days taken for germination, plant height
(cm) one month after germination of seed and survival percentage one month after germination of seed
for each pre treatment.
Totalgerminationinallreplicates
% ofGermination =
No. ofreplicates
The data collected for all the quantitative and qualitative traits was subjected to cluster analysis to
conduct similarity estimates using unweighted pair group method of arithmetic averages (UPGMA) in GGT.
Analyses were performed using the GGT 2.0 2010 software for Graphical GenoTypes.
1. Survey and collection of Buchnanania lanzan genotype (seeds and plantlets) from different Agro-
climatic zones of Chhattisgarh.
The experimental site, Chhattisgarh state stretches across the latitudinal expanse of 17°46' to 23°15'
North on one hand to the longitudinal meridian of 80°30' to 84°23' East on the other. The samples for the
present study were selected from all the three agro-climatic zones of Chhattisgarh shows in table 1. The
passport data of experimental sites recorded in table 2.
2. Morphophysio characterization:
Morphophysio data of fruits of all selected wild genotypes of Chironji in the period of may-june
from all agro climatic zones of Chhattisgarh in terms of fruit size, weight, colour and health of fruits,
number of fruits per kg, wt. of 100 seeds / kernel (obtained from 100 fruits). The results of morphological
attributes like Fruit size was found to be minimum (9 mm) in Jashpur region while the maximum fruit size
(12 mm) found Jagadalpur and Kondagaon region. Fruit colour changes gradually as growth development
and at different stages of maturity As the collection was done in second week of May the percentage of
black seeds in all locations were found to be maximum against Reddish green. Collection time period may
influences the health of fruits also as all the collected fruits found well ripped and healthy. Table 3.contain
the data for morphological attributes of selected genotype of Chironji
In Physiological analysis main emphasis was on the seed size, weight of seeds,% of purity, % of
Seed germination, and % Moisture of seeds. On the basis of statistical analysis of physiological
characterization data of all 27 samples of chironji genotype, Seed size was found to have increased level in
zone 1comparitively zone 2 and 3. Results for other attributes like Seed weight, purity Moisture and
germination % the results were best comparatively with other two zones.
The initial moisture content of Chironji nuts and kernels was found to vary from 6.00 to 9.00%
and from 2.00 to 4.00% (db), respectively. The weight of individual nuts varries from 0.17-0.40g. (Shilpa
Deshmukh et al., 2017).In present study the results found on percentage basis for moisture content while nut
weight results changes on basis of locations.
The statistical data for all the assessed characters of Chironji seeds are as follows in table no. 4.
All the genotypes were plateau stands out as most diverse from rest of the genotypes. divided into 2
major groups (A and B) based on DMRT analysis with 3 genotypes (Bb, Pp, and Md) in one group (A)
showing about 86% diversity between x, y and z and while x and y were 60% genetic similarity. Out of 27
Genotypes3 genotypes (Bb, Pp and Md of genotypes) showed 90.0% variability compared to other
genotypes. In particular, the genotype Md collected from Bastar
Genotypes Ms& Ap (from Raipur plains and Northern Hills region), Rt, Bh and Mg(from Raipur
plains, Bastar plateau and Northern hills region),Pr and Kt (from Raipur plains region),Gb, Ck, Mp ((from
Raipur plains, Bastar plateau and Northern hills region) and Sp, Pb, Kb(from Bastar plateau, Northern hills
and Raipur plains region) were 100% similar to each other on the basis of phenotypic characters. These
genotypes formed a separate sub-group within the ‘B’ group. While 9 genotypes sharing relatively lesser
genetic relatedness formed another subgroup, in which none were 100% similar. , genotypes Md collected
from Bastar plateau stand out from rest of the genotype.
Group A:
Three wild genotypes of Chironji from Bastar plateau agro climatic zones of Chhattisgarh clustered
in this group, these are Bastar accession accessions exemplified by large and medium-large sized fruits.
They are characteristically early and high yielding. Higher nut yield both in terms of number of nuts per tree
and the size of fruits. The potentials abound in this set of trees suggest possible selection of high yielding
varieties of Chironji that produce high quality grade nuts in the short term for the poor resource
Chhattisgarh tribal community. These materials will also be useful in Chironji improvement work.
Group B:
All remaining twenty four genotypes clustered together in this group. With other genotypes from all
the agro climatic zones sub clustered in five small clusters. The trees were having same growth patterns but
the A fruit yield in terms of fruits per tree was lower than Bastar Plateu. The difference in fruit size also,
they are somewhat small in size that Bastar region fruits. These materials would be good source of genes
that could be exploited in any improvement work to improve on fruit size and quality.
Fig 2. Showing all the steps A. Selection of superior wild genotype of Chironji. B .Flowering of
Chironji plant. C. Fresh and healthy fruits of Chironji. D. Seeds of Chironji E. Seed germination in
Green house F. Fully grown seedlings of Chironji
Graphical Representation
Seed weight
Seed Size SeedWeight
Seed weight % Purity
270
A 84.00%
0.415
v 265 82.00%
g s 0.41
Zone1 Avg Seed Zone1 % Purity 80.00% Zone 1
i 0.405 260
Zone 2 weight Zone2 78.00% Zone2
S z
0.4 255
e e Zone 3 Zone 3 76.00% Zone3
e 0.395 74.00%
250
d 0.39 Agroclimatic 72.00%
Agroclimatic zones
Zones
Agroclimatic zones
0.84% 94.00%
0.82% 92.00%
0.80% % of 90.00%
Zone 1 germinatio Zone 1
% of 0.78%
Zone2 n 88.00% Zone 2
Moisture 0.76%
Zone3 86.00% Zone 3
0.74%
84.00%
0.72%
82.00%
0.70% Agroclimatic Zones
Agroclimatic Zones
By Enlarge for all physiological traits Bastar Plateau (Zone 1) found to be superior
Graphical Representation
Bg
Gg
Gg
Bg
Kh
Km
Km
Km
Ph
Bb
Gb
Bh
Pp
Ck
Ap
Pb
Ph
Kb
Bb
Gb
Kh
Bh
Pp
Ck
Ap
Pb
Ph
Kb
Bb
Gb
Kh
Bh
Pp
Ck
Ap
Pb
Kb
Rt
Rt
Rt
%Moisture %Germination
1.40% 120.00%
1.20% 96.00%
100.00%
1.00%
80.00% 69.81%
0.80%
60.00%
0.60%
40.00%
0.40%
0.20% 20.00%
0.00% 0.00%
Km
Km
Bg
Ph
Bb
Gb
Gg
Kh
Bh
Pp
Ck
Pb
Kb
Bb
Gb
Gg
Kh
Bh
Pp
Ck
Bg
Pb
Ph
Kb
Rt
Rt
Ap
Ap
Genotypes Genotypes
The analysis of variance revealed that all traits showed large differences, indicating a high level of
morphological variation with two different groups between locations with 100% variability. According to
data and final results, Morphophysio analysis of all selected genotype of Chironji. In all tested samples of
chironji, Bastar plateau found to be superior in all characterization.
References: